Cocktail vaccine

Dr.M.Muruganandam

ISBN 978-9982-22-634-9

First Edition-2018

Author: Dr.M.Muruganandam.

Email: vaccine.m@gmail.com

Publisher

Einsteein Bio-Engineering

Research Foundation,

South India.

Preface

New strains of pathogens are

emerging day by day in the various parts of

the globe due to over population explosion and

manmade pollutants. These harmful materials

created genetic changes in the organisms as a

result, it will produced new strains of

pathogens. It will also create big burdens to

our society. For control and management of

this disease burden, we need multidirectional

combined approach of action. Vaccine

development is one of the control measures

which is simply explain in this book to young

readers. For the preparation of these

manuscripts, I have referred many

researchers’ work, I thank all of them.

M.Muruganandam

Content

Chapter 1.Vaccine

1.1 Immunogen

1.2 Efficiency of various Immunogens

1.3 Nucleotide Immunogen

1.4 Cocktail vaccine

Chapter 2. Current status

2.1 Peptide vaccine

2.2 Virus vaccine

2.3 Staph cocktail vaccine

2.4 Parasite vaccine

2.5 Cocktail DNA vaccine

2.6 Side effects

Chapter 3 Vaccine Development

3.1 Vaccine complex

3.2 Immunogen Mix

3.3 Bio-active compounds

Chapter .4 Problems to be solved

4.1 Children’s vaccine

4.2 Elder’s vaccine

4.3 Immuno suppressed

Patient’s vaccine

4.4 Food and water bone

Disease vaccine

4.5 Stress vaccine

4.6 Public Hygiene

Worker’s vaccine

4.7 Poor people’s vaccine

4.8 Hospital worker’s vaccine

4.9 Malarial vaccine

4.10 Multi drug resistant

Pathogens vaccine.

1. Vaccine

1.1 Immunogen

Vaccines save millions of

people from various infectious Diseases.

The vaccine has many important

compounds. The immunogen are one of

the important components of the vaccine,

which are able to stimulate cell mediated

and humoral mediated immune responses

in the host. However the efficacy of

various Immunogen is vary, but different

types of Immunogens are present in

pathogens. The important immunogen are

surface proteins, Toxic proteins,

nucleotides, polysaccrides, etc.

The vaccine immunity mainly

depends on the efficacy of its

Immunogens. During vaccine preparation,

if add more types of Immunogens leads to

develop new powerful cocktail vaccine. In

these type vaccines more types of

Immunogens used to increase efficacy.

Similarly different types of best adjutants

or mixer of best adjutants are used. It also

needs to boost up the efficacy of vaccine.

This is one type of cocktail vaccine.1

Another type of cocktail

vaccine also prepare for prevent more than

one disease. Here Immunogens of the

various pathogens are mixed and vaccine

will be developed. It will be used to solve

many problems in the growing population.

The cocktail vaccine-I is best candidate to

prevent multi drug resistant pathogens.

Because it is more powerful compare to

ordinary vaccine. It has combination of

different types of best Immunogens and

best adjuvant mixer, which will increase

the efficacy of vaccines.

Cocktail vaccine-II prevent

group of disease, such as food/water borne

disease, children’s diseases, public

Hygiene worker’s, vaccine, Hospital

worker’s, stress vaccine, etc. So this type

of vaccine saves population at risk

condition.

1.2 Efficacy of Various Immunogens

Antigenic efficacy of bacterial

Immunogens is varying. The qualitative

and quantitative levels of these

Immunogens determine the efficacy

depends on many internal and external

factors. The important factors are dietary

composition (Nutritional factors)

conditions of the immune system, Amount

of specific Immunogens in vaccine,

different types of Immunogens in vaccine,

selection of adjuvant for vaccine

development, etc. However in our lab

trial, the efficacy of immunogens was

studied in bacterial pathogen, Aeromonas

hydrophila.

Aeromonas hydrophila

is a motile rod shaped bacterium. The

Aeromonas sp. are water borne

microorganism that have seen implicated

reputedly as the causative agents of

clinical illnesses of serious, ranging from

gastro -intestinal and wound infections to

septicemia .2 Bacteraemia is the most

common pathogenic manifestation of

Aeromonas. In humans its mild symptoms

include fever and chills but patients who

become septic with A.hydrophila often

exhibit abdominal pain, Nausea vomiting

and diarrhoea.

Vaccine as an antigenic

preparation used to produce active

immunity to a disease in order to prevent

the effects of infection by any natural or

wild strain of the organism. Vaccine

prepares our body for fight deadly disease

by triggering of persons immune system,

this leads to the production of immune

response that can fight to the germs. The

vaccine contains a killed or weak end

form or derivatives of the infectious

germs. But recent advances in molecular

biology have provided various methods

for producing vaccines. But still

researches are going on to find out proper

vaccine for the Aeromonas infectious

disease in the world wide. Now there is no

vaccine in Aeromonas hydrophila

infection due to its antigenic diversity. In

this study, various Immunogens are

isolated and tested their efficacy for

vaccine development against Aeromonas

hydrophila infection.

The Aeromonas

hydrophila (MTCC – 646) strain was used

for this experiment Albino rats were used

as test animal in the whole study. Albino

rats were purchased from local animal

husbandry and they were acclimatized to

the laboratory environment for a week.

They were maintained in cool dry place.

During this study, the room temperature

ranges from 26-29oC, Daily they were fed

on commercially available feed and

inspected daily.

In this study, two

experimental trials were conducted. In the

first experiment, four set of albino rats

were used each set contain three rats being

exposed to killed live attenuated and

toxoid vaccine. In the second trial

Genomic DNA, Plasmid DNA and whole

protein were used as vaccine. Saline was

used as control in both experiments. After

15 days pathogens were infected and

primary immune response were studied.

During killed vaccine

preparation approximately 1ml of nutrient

broth inclusive of Aeromonas hydrophila

was centrifuged at 10,000 rpm. for l0

minutes and the pellets were collected and

removed the supernatant then saline was

added to the pellet and it was heated in

boiling water (30 minutes) followed by

centrifugation at 10,000 rpm and the dead

cells were collected. It was serially diluted

at 10-5 with saline.

For live alternated

vaccine preparation, the cells were

isolated first from the old culture in

nutrient medium and then these were

serially diluted to 10-5 using saline. For

toxoid vaccine preparation, viable calls

were inoculated in to nutrient broth and

inoculated for 24 hours. The content was

centrifuged at 10,000 rpm for 10 minutes.

The supernatant was collected and serially

diluted to using saline.

During DNA vaccine

preparation, first genomic DNA and

plasmid DNA were isolated from bacterial

pathogen. For whole protein vaccine

preparation, Ammonium sulphate was

used. After isolation of proteins, it was

serially diluted up to 105 by using saline.

For Hematological analysis, regular

procedure was followed. For antibody

analysis, 96 well microtitre plate was used

and also immuno electrophoreses was

carried out.

In the results of first

experiment shows that, maximum WBC

count (9200 cells,/cumm) was observed in

killed vaccine treatment other parameters

such Hemoglobin (12.2gm%) RBC counts

(4.06 cells /millions) and PCV (36.6) were

lesser compared to toxoid and live

attempted treatments. The second higher

value of WBC count (8200 cell/ cu.mm)

was observed in live attenuated treatment.

The control treatments have lesser value

compares to other treatments. The

maximum hemoglobin (13.gm%) and

RBC count (4.36 cells/Millions)were

observed in toxoid treatment.

The higher antibody

production was observed in killed and live

attenuated treatment compared to toxoid

and control treatment. So in the first

experiment, In activated (killed) vaccine

has maximum immune response compared

to other Vaccine treatment.

In the second

experiment, higher value of WBC counts

and antibody production was observed in

protein and plasmid DNA treatment. In

the counter current immuno

electrophoresis results shows that more

than one type of antibody was produced in

plasmid DNA treatment compared to

other vaccine treatment.

In each part of the

pathogen has various level of

immunogenic efficacy. The increased

level of some components and appropriate

level of mixer of various immunogen

determine the immunogenic efficacy of

the vaccine. In this trial various

components of bacterial pathogen

(Aeromonas hydrophila) was isolated and

tested their immunogenic efficacy in

albino rats. The conventional approach to

the process of vaccine development

includes the use of inactivated whole cells

and live attenuates microbes.

The major drawback

with these two approaches is that large

quantities of organisms are usually

required to isolate sufficient antigens for

prepare a vaccine. The modern approaches

of vaccine developments are best

compared to old approaches. Another

important drawback is in killed vaccine, it

cannot multiply in the host. So the levels

of antigens are always same at long time

but live attenuated vaccine continue to

multiply inside to the host. So the

immunity may increase and present in

long term. Recently it has been increasing

interest that the use of live attenuated

vaccines against bacterial pathogens.

In general, live vaccines elicit a

stronger cell-mediated response then

bacteria 3 While the greater immunity

provided by attenuated organisms

compared with that provided by dead

bacteria may be explained by induced

expression of stress proteins and possibly

certain abundant toxins, Aeromonas

within the host1.

In the second trial three

types of test vaccines are used. One is

genomic DNA another one is plasmid

DNA and third one is whole cell protein.

The DNA vaccine is under trial in various

laboratories, which is being tested in

human beings, the injection of plasmid

loops of DNA that contains genes for

proteins produced by the organisms being

targeted for immunity. In this study

plasmid DNA treatment produce more

antibodies against A.hydrophila compared

to genomic DNA. Similar trial was done

in staphylococcus aureus, salmonella

typhi and E. coli.4

In the first trial live

attenuated vaccine produce more

immunity. In the second trial protein and

plasmid DNA produce higher immune

responses. Usually the surface antigenic

immunogens are proteins which is

produce higher immunity .If use mixer of

protein and plasmid DNA immunogens

the efficacy of vaccine will reach high. So

this preparation is the recommended to

New Aeromonas hydrophila vaccine

preparation.

1.3 Nucleotide Immunogen

The immunogen are important

components of the vaccine which is able

to determine the efficacy of vaccine. It is

also immunity and immunological

memory. The nucleotides are also act as

immunogen. The bacterial plasmid DNA

has few nucleotides. Nowadays multi drug

resistant pathogens appear in many

countries. It gives global threat WHO

advices to produce new vaccine against

these short sequence plasmid DNA opens

new windows to develop novel vaccine.

Table -1. Immunogenic efficacy of various cellular

components of Aeromonas hydrophila.

S.no

A B C D

–L

D-

P

E F G

1. Control 3.9 7,500 15 85 11.8 38.25 07

2. Killed

Vaccine

4.05

9,200 23 75 12.2 38.75 10

3. Live

Attinated

Vaccine

4.25

8.900

25

72

12.8

39.75

09

4. Toxoid

Vaccine

4.25

8,800

12

88

13.1 36.75 08

5. Protein

Vaccine

4.3

9,200

18

81

13 39.75 09

6. Genomic

DNA.vac

4.33

8,700

11

86

12.8

37.25

08

7. Plasmid

DNA.vaccine

4.28

8,000

20

79

13.1

37.25

08

A=Cellular component, B=R.B.C count ,C=W.B.C total

counts, D=W.B.C Diff.counts, P(%),L(%) E=Hb(gm),

F=PCV,G=Antibody titre .

For more than one disease

prevention, cocktail vaccine trials are

conducted. During our lab trials, the

maximum immune responses were

observed in mixer of digestive plasmid

DNA compared to undigested plasmid

DNA mixture.10 during optimization trials

of S.typhi oral plasmid DNA vaccine the

maximum efficacy was observed in

double digested plasmid DNA vaccine

compared to whole plasmid DNA

vaccine.5

1.4 Cocktail Vaccine

It is a mixer of more

than one species of same pathogen or

different pathogen. The aim of the

cocktail vaccine development is to prevent

the spreading of multi-drug resistant

pathogens and also prevent the group of

diseases in specific population In our

study, Still many parasites gives headache

to scientist during vaccine development.

Cocktail vaccine development solve this

problem and it well help to develop best

vaccine against parasites. The antibiotic

resistant staphylococcus aureus bacterial

pathogens produce more problems during

vaccine development. It is mainly due to

presence of more strains and also multi-

drug resistant power. This cocktail

vaccine gives new scope to vaccine

development. Because immunogen of

various strains are used in cocktail

vaccine. The design and preparation

methods are easy and it can be alter or

adjusts according to practical need. In

future the cocktail vaccine help to solve

many common disease problems and save

human society from various risk

conditions.

2. Current Status

2.1 Peptide vaccine

There are different types of

antigenic peptide present in bacterial

pathogens, their immunogenic efficacy

also vary. The important immunogenic

peptides are surface proteins, Heat stress

proteins, Toxoid proteins, etc. These are

used to develop vaccine for single

pathogen or sometimes these mixtures of

peptides help to prepare a vaccine to

control more than one diseases. Based on

the requirement, the cocktail vaccine will

be altered.

During cocktail vaccine

preparation appropriate amount of

different types of immunogenic peptide

mixed and vaccine will be prepared. Each

peptide is prepared separately and

combined in to a single drug product.

Nowadays the multiple peptides mixed

and single drug product developed, this is

also a good approach for development of

vaccine.

2.2 Virus Vaccine

The virus cocktail vaccine is under

the experimental trials. Now scientists are

making the case that a vaccine against

rhinoviruses, the predominant cause of

common cold is achievable. The vaccine

against rhino viruses may be questionable,

because there are more than 100 varieties

circuiting around the world. It’s

supporting that nobody tried such a simple

solution over the last 50 years. We just

took 50 types of shini virus and select

dominant type then mixed them together

into our vaccine.

A mixture of different types of in

activated rhinovirus stimulate neutralizing

antibodies performs well against all the

viruses in mice. Similar things are also

repeated in rhesus macaques. In this work,

antibodies generated in response to the

vaccine were tested and analyzed for their

ability to prevent the virus from infecting

human cells in culture.1

2.3 Staph cocktail vaccine

Antibiotic resistant Staphylococcus

aureus is one of the global threats.

Different strains of staphylococcus aureus

cause disease, for solve this problem

mixer of Immunogen from different stains

are used. In the same time immunogens of

more virulent pathogens are mixed and

vaccine will be developed. This type of

cocktail vaccine in Staphylococcus aureus

was prepares and tested in albino rats. It

produces good results2 .Now research

work is going on to develop effective

vaccine against staph infection.

2.4 Parasite Vaccine

The parasite life cycle has different

complicated stages. So still it is very

difficult to develop vaccine in many

parasitic diseases. But cocktail vaccine

gives solution to parasitic diseases. The

mixture of immunogens from various

parasitic life stages are isolated, identified

and mixed appropriate levels and based on

the requirement, vaccines will be

developed. The parasite vaccine

development is under trial level because

the host parasite immunological

relationship is not completely understood.

Much parasite vaccines are still under

experimental levels, due to presence of lot

of hurdles and complications in vaccine

development. For eg. Malaria vaccine,

Filariasis vaccine etc,

2.5 Cocktail plasmid DNA Vaccine

The cocktail DNA Vaccine is also

under research level. In our lab trials two

types of cocktail DNA Vaccine were

developed. One is plasmid DNA Vaccine

and another is genomic DNA vaccine. The

details are explained one by one. First

plasmid DNA Vaccine was developed for

various bacterial pathogens. The digested

plasmid DNA was also used.3 In this

work, mixer of three different pathogen’s

plasmid DNA based cocktail vaccine was

developed.

In these studies three bacterial

pathogens such as salmonella typhi,

staphylococcus aureus and Escherichia

coli were collected from local hospital

laboratory. All the biochemical and micro

biological examination were done for

confirmation. Then prepared three

separate broths and individually

inoculated. After 24 hours plasmid DNA

was isolated separately from culture and it

was isolated by alkaline layers method. In

the first treatment all the plasmid DNA

were mixed and used as vaccine.

In the second treatment all the

plasmid DNA were digested by Bam H.1

restriction Enzyme. Then it was used as

vaccine. In the third treatment plasmid

DNA was isolated from different culture

and digested by PST-1 restriction enzyme

and there is mixed then used as vaccine.

In the fourth treatment, plasmid DNA was

isolated from all the pathogens and double

digested by using Bam H-1+pst-1

enzymes and they were mixed well then

used as vaccine. One control treatment

was always maintained throughout the

experiment. Albino rats were used as test

animals in all the treatments. After

delivered the test vaccines two weeks later

blood samples were collected for analysis.

In these study maximum immune

responses was observed in double

digested plasmid DNA treatment

compared to other treatments. The second

maximum immune response was observed

in undigested plasmid DNA treatment.

The lesser response was observed in

control treatment restriction enzymes Pst-

1 digested plasmid DNA treatments gives

better results than Bam H-I digested

treatment.

The RBC count was not much

charge in all the treatments. It was more

or less some level in undigested plasmid

DNA treatment. Pst -1and Bam H-I

digested plasmid DNA have similar RBC

counts. Higher level of polymorph and

lymphocytes were observed in double

digested plasmid DNA treatment,

compared to other treatments Antibody

levels were higher in single and double

digested treatments. The control treatment

has lesser antibody levels compared to

other treatment.

2.6 Genomic Cocktail Vaccine

The another type of cocktail

vaccine is Genomic cocktail DNA

vaccine..In this study, three pathogens

(Salmonella typhi, Staphylococcus aureus

and, E.coli) were used. First genomic

DNA was separately isolated from these

three pathogens. In this study three

treatments were tested. In the first

treatment, the entire DNA were mixed and

used as vaccine. In the second treatment

the DNA was digested by bamH-1 and

mix well than used as vaccine. In the third

treatment DNA of all the organisms was

digested by PST -1 enzyme then mixed

well.

Table : 2 Cocktail Plasmid DNA and Genomic DNA vaccine influence on

immune responses in Albino rats.

Treatm

ents

WBCCo

unts

(tells/in

mm)

W

BC

P

Diff.

Coun

t(L

%)

(

%

)

E

RBC

Count(mi

i-

llionns/cu

mm)

Hb(

%)

Antid

ody

Titre

A 4800 20 78 02 3.7 14.5 7

B 7100 30 70 -- 3.3 9.9 8

C 5900 27 65 03 4.2 12.5 10

D 6600 19 81 -- 4.3 13.0 10

E 7600 22 78 01 3.5 12.3 10

F 5500 18 62 -- 3.8 11.5 7

G 4900 31 68 01 3.5 10.6 10

H 6300 24 70 01 3.5 10.6 10

A=control, B=Whole plasmid DNA mix,

C=Bam H I digested Plasmid DNA mix,

D=PST-I digested plasmid DNA mix,

E=Doubled digested Plasmid DNA mix,

F=Genomic DNA mix

G= Bam H-I digested genomic DNA mix

H=PST-I digested genomic DNA mix

and used as vaccine. One control

treatment was also maintained.

After one week of injection same

booster dose was given to all the

treatments. At the end of the experiments,

blood samples were collected for analysis.

The maximum RBC count was observed

in PSt-1 treatment and the minimum WBC

counts were observed in control treatment.

The highest antibody level was observed

in enzyme digested DNA treatment. So it

is concluded that PSt-1 digested mixer

genomic DNA acts good vaccine

compared to other cocktail genomic DNA

vaccine.

The Correct design and appropriate

combination of all the components are

essential in cocktail vaccines. If the

composition is not correct it may leads to

inefficient functions of vaccines. In the

first gulf war, cocktail of Vaccines

administrated to troops, some are received

as many as 12 vaccines at one time and

went to develop severe reactions.

The cocktail of inoculations lead

them to suffer from symptoms similar to

those for chronic fatigue. They

complained of headaches, memory loss,

muscle pain, nausea, gastrointestinal

problems, loss of concentrations, vision

and balance problems. People claims that

many hundreds of former military

personal have died from gulf war

syndrome. This is happened due to

improper study of synergic reactions of

vaccine. After the clear study, only the

cocktail vaccine should introduce. If

introduce without clear study leads to

develop many problems.

3. Vaccine Development

3.1 Vaccine complex

Cocktail vaccine contains more

than one immunogen from one pathogen

or different strains/types of the pathogens.

Based on the requirement, the vaccine will

be designed. The important components is

of the cocktail vaccine are as follows.

• Immunogen mix,

• Bio-adjuvant

• Bio-Preservatives and

• Bio-active compounds

First different components are

standardized and mixed appropriate

amount, then vaccine will be developed.

These components are discussed one by

one.

3.2 Immunogen mix

The immunogens are mainly

determined the efficacy of vaccine. There

are different types of immunogens used in

different vaccines. In the cocktail vaccine,

mixer of all the immunogen are used such

as inactivated pathogens, various

antigenic proteins and fragments of DNA,

etc. The inactivated cells induce

immediate immunological reactions and

immunological memory. But the numbers

of cells are not multiply, The number is

constant, so it cannot produce long term

immunity, the immunity will be fade

slowly. But for short term immunity, it is

good because it produce higher level of

immunity.

For inactivated cell preparation,

there are three methods are followed such

as formalin in activated cells, Heat killed

cells and antibiotic in activated cells. In

the first method, cells are harvested from

broth through centrifugation then 0.5% of

formalin will be added and incubate at

4oC in overnight. After that all the cells

are washed with the help of physiological

saline. Now it is ready to use.

The second method is in activated

by heat. The bacterial cells are introduced

in 1000C water bath for 30minutes then

these cells are used in vaccine. The third

method is using suitable antibiotics to kill

the bacterial cells. After killed the

bacterial cells, they are suspended in

physiological saline. These materials are

stored for vaccine preparation and are

portion of these materials is tested. They

were introduce in spread plate then it is

incubate 370c for overnight or

24hours,after that check, if any live cells

are present in solution or not, then

confirmation of all the cells are at death

condition, they were serially diluted and

used in vaccine preparation.

Second thing is Antigenic protein

preparations. Different types of antigenic

proteins present in the bacterial cells. The

important antigenic proteins are surface

protons, toxoid proteins, Head Stress

proteins, etc.. During extra cellular proton

preparation, the cells are removed from

the culture by centrifugation or using

appropriate filter cells are filtered then

protons are precipitated and also

inactivated. The osmoregulation is also

help to isolate the surface proteins in

pathogens. If use hypertonic/hypotonic

medium, it can easily remove surface

region of the cell, using Various methods

different types of antigen proteins are

isolated, optimized and finally all are

standardized then they will be used in

Vaccine preparation. In the case of

preservatives , compared to artificial

chemicals, Natural product based

preservatives are good, cheap, possible to

receive lesser side effects and also

highly potential compounds .All types of

Natural preservatives are not still

exposed. Many natural compounds,

antimicrobial peptides present in marine

sources. In future, it may expose and new

Compounds will be use as preservatives in

vaccine development. Based on their

efficacy and functions, it will use as single

Compound or mixture of Compounds will

be used as bioperservatives in vaccine

development.

In the Animals sources

lot of anti-microbial peptides are present.

still researchers are needed to explore

these peptides. In our lab trial,

antimicrobial compounds observed in fish

internal organs. In this study, various

internal organs of fish was isolated and

prepared extract with the help of

chloroform and ethanol (1;1) mixture

Antibiotic disc were pre pared by using

these extracts. They were prepared by

using these extracts. They were placed on

the bacterial pathogen (Aeromonas

hydrophila) spread plate, after

appropriate time incubation, Zone of

inhibitions were observed.

The maximum Zone of

inhibition is observed in Brain and muscle

tissue extracts. So it is concluded that, fish

internal organs contain some antimicrobial

peptides and proteins. The discoveries of

various Natural antimicrobial compounds

gives new s scope to develop new

preservatives in advanced vaccine

development .New researches in various

marine animals are must to discover new

Compounds. In Marine environment still

many unknown drugs and antimicrobial

compounds are present and it is good

resources to solve many essential

problems in our society.

3.3 Bio-active compounds

In therapeutic vaccine development,

Bio-active Compounds are added during

vaccine development. The therapeutic

vaccine used in after infectious

Condition. So it needs to reduce the

pathogen load. It Contain mainly bio

active Compounds and the remaining

vaccine Compounds which trigger the

immune system against infection.

The marine resources contain

varieties of bio-active Compounds.

However these varieties of bio-active

Compounds are not yet used in vaccine

development. Still more and more

research work are need in this area The

main function of therapeutic cocktail

vaccine Components strengthen the

immune system and it help to fight

against the pathogens. The Natural bio-

active Compounds are healthy, low cost,

easily available, lesser side effects

Compared to Synthetic one.

4. Problems to be solved

Cocktail vaccine helps to prevent

group of infections. It can be easily solve

many problems in our society. In different

condition, more than one pathogen is

involved in the infection. In chronic

diseases condition, many bacteria and

fungi are involved the secondary

infections. If develop cocktail vaccine, it

will help to solve these problems and it is

also helped to control multidrug resistant

pathogens.

4.1 Children vaccine

Children have weak

immune system and the immune system is

not fully developed. They cannot fight

against many infectious disease at a time

for example diseases in raining season or

some other seasons, at that time suitable

cocktail vaccine gives protection from the

diseases. The therapeutic vaccine is

needed to maintain children’s health,

because it will use after infections, for this

vaccine development, screening of broad

spectrum medicine is first step. If develop

a cocktail vaccine, it will prevent many

infectious diseases at a single dose of

medicine in early childhood condition.

4.2 Elder’s Vaccine

Aged populations immune

system is also weak. They suffer from

various age related diseases. So it needs to

develop suitable cocktail vaccine for those

diseases. If develop therapeutic cocktail

vaccine, it reduce disease burden and

induce functions of immune system and

also gives strength to the immune system.

In the therapeutic vaccine development

drugs, it will reduce the side effects and

save people’s health.

4.3 Immuno-suppressed patient’s

Vaccine

The immuno- compromised people

have impaired immune system. They need

special design for vaccine development,

because the new vaccine will give more

strength to immune system and induce to

fight against the infectious diseases. If add

bio-active Compound in the vaccine, it

will help to clear the pathogen load and

reduce burden of the disease. The diet

plays an important role in the functions of

immune system. Many nutritional factory

acts as anti-oxidants and immuno

stimulants. They help to strengthen to our

immune system. If design table of diet

Components and rations it will help to

improve the functions of immune system

and also functions of vaccine. The

nutritional immunology plays a important

role in vaccination of immuno-

compromised patients.

4.4 Food and water borne Disease

vaccine.

In developing Countries, very

difficult to provide safe food and drinking

water to all the people. In most of the

traditional celebrations, religious

functions the water borne diseases are

more chance to be explored and

sometimes created major problems

because people assemble in one place

during the celebrations. At that people

cannot maintain good hygiene and

difficult to maintain in safe drinking

water. In rural areas and also most of the

town and cities in developing Countries,

people maintain poor sanitation and public

hygiene. So the drinking water bodies are

unprotected and also possible to pollute.

The Contaminated water Contain many

diseases, it will possible to explore in that

area. So in this Condition Cocktail

vaccine is essential to save people form

group of water borne diseases.

In poor Countries, small

hotels with various food varieties present

in everywhere but the level of hygiene is

very low they use low quality ingredients

and also use poor hygiene drinking water

and other materials. The food preparation

Conditions and place have to maintain

very poor sanitations, it leads to possible

to spread more infections disease through

contaminated food and water. In future If

develop therapeutic cocktail vaccine, it is

good because it reduce disease burden and

also educate to our immunesystem to fight

against various infectious diseases.

4.5 Stress vaccine.

In the modern life style

provide more stress to people. The target

oriented western life cycle creates more

stress and it induces to produce more

cortisol hormones which lead to provide

more stress and stress related problems. In

day today life, all the people affected in

various type of stress, mostly affected by

mental stress. Physiologically, it creates

lot of problems which aggravate other

diseases. In this condition there is need to

stress reducer therapeutic cocktail vaccine

and common food schedules for reducing

stress and save the people. More

researches are need in this area to develop

cocktail vaccine which is suitable for all

types of people.

4.6 Public Hygiene worker’s Vaccine. In

most of the developing courtiers, the

public sanitary workers and Hospital

cleaning workers are affected forms

various infections disease. Suppose if

prepare common therapeutic vaccine, it

will help to Control these types of

problems and save the public workers

health Condition. Mostly sewage cleaning

workers, hospital cleaning workers and

Nurses are victim for many infectious

diseases. So the cocktail Vaccine

development gives better protection to this

group of infectious diseases. More and

more research works are needed in this

area to develop various cocktail vaccines

in developing Countries.

4.7 poor people Vaccine.

In the day today life of villagers,

they spend most of the time in Agriculture

filed. They are mostly work in soil and

also spend in animal husbandry. During

their work , they may be infected by

various bacterial, fungal and parasitic

pathogens. The cocktail prophylactic

vaccine and therapeutic Vaccine provide

solution for all these problems. It will

protect common infections. More and

more research work is needed to develop

cocktail vaccine to solve these problems.

In the city and Town, poor people are

mostly suffer in malnutrition and poverty

related diseases, For this situation, the

cocktail Vaccine is need to save the

people from those disease condition.

4.8 Hospital worker’s Vaccine

In the hospital atmosphere, most of

the pathogens are drug resistant. If people

stay long time or working in long time,

they may get more chance to various

infections. The hospital workers have

more chance to get infections form

multidrug resistant pathogens. So they

should follow proper precautions

methods, otherwise they may suffer in

various infections in long term. The

cocktail vaccine helps to solve the

problem. It gives protection form these

diseases and gives more strength to our

immune system, the discovery of

Common cocktail vaccine for hospital

workers is very important because they

save people.

4.9 Malaria Vaccine.

In malaria the host –parasite

interaction at molecular level is not

completely known. If understand these

mechanism which help to develop new

vaccine. The second thing is life style of

parasite has many complicated stages. So

Identification of correct Immunogens are

very big problem. Now drug resistant

malaria reported in some places.

However, the cocktail vaccine enhances

the efficacy. If it is also to help fight

against these pathogens. Most of the

developing Countries are suffering in

parasitic infections and drug resistant

pathogen problems. Because very

difficult to change life style and economic

status of people. This cocktail Vaccine

development is helpful to control these

type of problems in future.

4.10 Multidrug resistant pathogens

vaccine.

The multidrug resistant pathogens

populations is slowly increase in all over

the world especially in developing

Countries The main reason is misuse of

antibiotics and over use of antibiotics. For

Control these things, we need new drugs

form nature especially from marine

resources. The marine environment has a

lot of unexplored compounds and drugs.

More researches are needed to discover

best drugs. Nowadays antimicrobial

peptides discovered in many animals and

humans, these are also helpful to control

pathogens.

The new therapeutic cocktail

Vaccines are under research Condition In

future good therapeutic Vaccine help to

manage the multidrug resistant pathogen

problem. The developing countries mainly

suffer in multidrug resistant pathogens

infections. Because there is more

population lesser hygiene, poor life status

in developing Countries and they have

weak economic states. So they are suffer

in research and drug development anyhow

I hope cocktail vaccine solve poor people

infections disease in future.

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About Author…

Dr.M.Muruganandam is a Scientist

in Einsteein Bio-Engineering research

foundation, South India. He published more

than hundred publications. He has an

Editorship in African Journal of

Biotechnology, International Journal of

Medicine and Biomedical Research, Journal

of Medicine and Medical Research, London.